Identifying Determinants of Borrelia burgdorferi and Peromyscus leucopus symbiosis

项目摘要/摘要： 我的博士后研究重点是了解莱姆病的病原体伯氏疏螺旋体是如何 在自然界中生存-这涉及到研究它与自然哺乳动物水库宿主的关系， 白斑拟青霉。虽然先前的研究表明，肩部硬蜱不成比例地跳动 通过喂食无症状的白纹伊蚊小鼠来获得伯氏杆菌(然后可以传染给人类)，它 目前尚不清楚(A)为什么白纹伊蚊似乎比其他小型啮齿动物更适合传播伯氏杆菌，(B) 为什么白纹伊蚊在慢性系统性伯氏杆菌感染后仍无症状？(C)是什么分子？ 伯氏杆菌用来成功感染白纹伊蚊的工具。勃氏假单胞菌的自然生态研究 代表着一个重要的公共卫生目标，因为，虽然伯氏杆菌感染通常是可以治疗的，但大量的 部分患者会在几个月到几十年内保持症状。因此，找到预防B。 伯格多费菌感染是严重的。这个项目的长期目标是更好地理解 伯氏杆菌与白纹伊蚊的关系揭示破坏地方病循环的靶标 减轻莱姆病的负担。 这笔赠款旨在更好地了解白头翁-B。伯氏杆菌宿主与微生物的相互作用：(A) 监测伯氏杆菌感染和白念珠菌炎症变化的实验工作 时间，以及(B)确定白念珠菌感染所需的伯氏杆菌基因。为了第一个目标，我会 试图了解白假单胞菌组织中的细菌负荷如何与组织病理学、扁虱感染性、 以及基因表达随时间的变化。这将涉及到对B。 到目前为止在这些啮齿动物中感染伯氏杆菌，并将检验各种假设，这些假设已经被假设，但 没有经过实地测试。为了做到这一点，我将利用数字液滴聚合酶链式反应，核糖核酸测序，显微镜 组织学分析和异种诊断测试。值得注意的是，我将同时执行相同的实验 两个小鼠自交系(C57BL/6J和C3H/母鸡)表现出轻、重度莱姆病样 症状)，以将我的发现与常用的小鼠模型进行比较。对于第二个目标，我 我将使用我的实验室过去经常使用的突变体库进行转座子测序筛选 出版物。总之，这些实验将极大地提高我们对宿主和 促进伯氏杆菌在自然界生存的致病因素。此外，这个项目非常适合推广 我的长期目标是在一家学术机构启动一个独立的研究项目，作为数据集 这项研究产生的结果将作为我未来实验室的基础。

Project Summary/Abstract: My postdoctoral studies focus on understanding how Borrelia burgdorferi, the causal agent of Lyme disease, survives in nature—which involves studying its relationship with its natural mammalian reservoir host, Peromyscus leucopus. While previous work has demonstrated that Ixodes scapularis ticks disproportionately obtain B. burgdorferi (which can then be passed onto humans) by feeding on asymptomatic P. leucopus mice, it remains unclear (a) why P. leucopus appear better suited than other small rodents to pass on B. burgdorferi, (b) why P. leucopus remain asymptomatic despite chronic, systemic B. burgdorferi infection, and (c) what molecular tools B. burgdorferi uses to successfully infect P. leucopus. Elucidating the natural ecology of B. burgdorferi represents an important public health goal, as, while B. burgdorferi infection is typically treatable, a substantial fraction of patients remains symptomatic for months to decades. Thus, finding mechanisms to prevent B. burgdorferi infection is critical. The long-term goal of this project is to better understand the symbiotic relationship between B. burgdorferi and P. leucopus to reveal targets for disrupting the enzootic cycle and reducing the burden of Lyme disease. This grant seeks to better understand the P. leucopus-B. burgdorferi host-microbe interaction by (a) performing experimental work to monitor the dynamics of B. burgdorferi infection and P. leucopus inflammation change over time, and (b) identify B. burgdorferi genes that are required for P. leucopus infection. For the first goal, I will attempt to understand how bacterial burden in P. leucopus tissues correlates with histopathology, tick infectivity, and changes in gene expression over time. This will involve performing the most comprehensive study of B. burgdorferi infection in these rodents to date and will test a variety of hypotheses that have been postulated but left untested in the field. To accomplish this, I will leverage digital-droplet PCR, RNA-sequencing, microscopic histology analyses, and xenodiagnostic testing. Of note, I will simultaneously perform the same experiments in two Mus musculus inbred lines (C57BL/6J and C3H/HeN, which develop mild and severe Lyme disease-like symptoms, accordingly) to compare my findings against frequently used murine models. For the second aim, I will deploy transposon sequencing screening using a mutant library that my lab has frequently utilized in past publications. In sum, these experiments will dramatically improve our understanding of both the host and pathogen factors that facilitate B. burgdorferi survival in nature. Additionally, this project is well-suited to promote my long-term goal of starting an independent research program at an academic institution, as the datasets generated in this study will serve as the foundation of my future laboratory.